Hi all
I currently use the MiSeq for 16s metagenomics using the V3-V4 primers from Caporaso et al (2010). I've finally nailed down library quantification and optimal cluster generation, but I've been having issues with sample evenness.
Currently I'm using PicoGreen to quantify my samples individually and then pooling equimolarly for qPCR. The range for sampling depth on my samples (~150 per run) ranges from 80k-350k. While I'm happy to be getting great quality data, I'd like to nail down the eveness on the MiSeq.
Anyone have any suggestion for optimal evenness?
Thanks,
Pat
I currently use the MiSeq for 16s metagenomics using the V3-V4 primers from Caporaso et al (2010). I've finally nailed down library quantification and optimal cluster generation, but I've been having issues with sample evenness.
Currently I'm using PicoGreen to quantify my samples individually and then pooling equimolarly for qPCR. The range for sampling depth on my samples (~150 per run) ranges from 80k-350k. While I'm happy to be getting great quality data, I'd like to nail down the eveness on the MiSeq.
Anyone have any suggestion for optimal evenness?
Thanks,
Pat
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